• Journal of Applied Biological Chemistry
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• v.66
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• pp.7-14
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• 2023

The effects of fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver extracts (FAAE) on regulation of inflammation and apoptosis were investigated in primary cultured rat cartilage cells. To identify the protective effects of FAAE against H₂O₂, cell survival was measured by MTT assay. Smad3, Collagen type I, MMP3, and MMP13 were measured by real-timpe PCR and westernbot and the inflammatory (NF-κB pathway, COX-2, iNOS) factors were determined by western blot. The apoptosis related factors (JNK, c-Fos, c-Jun, caspase 3, Bax, and Bcl-2) were determined by western blot. FAAE significantly increased the follwing: H₂O₂ treated cell survival, mRNA and protein expression of Smad 3, collagen type I. In addition, FAAE significantly decreased the protein expression of inflammatory and apoptosis related factors. This study suggests that FAAE have a protection effect of chondrocytes through inhibition of inflammation and apoptosis. Thus, FAAE is a therapeutic potential food componet in osteoarthritis.

• Applied Chemistry for Engineering
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• v.30 no.4
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• pp.479-486
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• 2019

This study investigated the anti-inflammatory effects of mixed extracts of Achyranthes japonica Nakai (Aj) and Aralia continentalis Kitagawa (Ac) (ratios of 1 : 2, 1 : 3, 1 : 5, 2 : 1, 3 : 1 and 5 : 1) on RAW264.7 macrophages. Cell toxicity was determined using a cell counting kit (CCK) assay. We evaluated anti-inflammatory effects of the mixed extracts of Aj and Ac by measuring interleukin $(IL)-1{\beta}$, IL-6, and tumor necrosis factor $(TNF){\alpha}$ using an enzyme-linked immunosorbent assay (ELISA) kit assay. The mixed extracts of Aj and Ac inhibited lipopolysaccharide (LPS)-induced $IL-1{\beta}$ and $TNF{\alpha}$ in LPS-stimulated macrophages. Comparing different ratios of the mixed extracts, the 2 : 1 ratio of Aj and Ac has much more potency and inhibited the production of $TNF{\alpha}$ in LPS-induced RAW264.7 cells. The results of the present study showed that the mixed extracts of Aj and Ac have potential anti-inflammatory effects on RAW264.7 macrophages. Therefore, these extracts may be used as a good source of functional foods for the protection against inflammatory diseases.

• Applied Chemistry for Engineering
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• v.30 no.2
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• pp.145-150
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• 2019

In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of $100{\mu}g/mL$, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and Interleukin-$1{\beta}$ (IL-$1{\beta}$), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.